Effects of Lin28a and Lin28b on let-7 family activity / 病毒学报

In this report, we study the effects of over-expression of Lin28a and Lin28b on let-7 family activity in HeLaS3. Firstly, we constructed pAAV2neo-Lin28a and pAAV2neo-Lin28b to express Lin28a and Lin28b, respectively. Then, pAAV2neo-Lin28a and pAAV2neo-Lin28b were transfected into HeLaS3, selected with G418 and obtained cell lines, HeLaS3/pAAV2neo-Lin28a and HeLaS3/pAAV2neo-Lin28b, to express Lin28a and Lin28b stably. Thereafter, we constructed eight plasmid vectors for detection of let-7 family activity based on pAAV2neo-Gluc-(Fluc). These vectors were further packaged into recombinant adeno-associated viral vectors (rAAV) which were used as sensors, nominated as Asensors, to detect inhibition activity of miRNA at post-transcriptional level. Subsequently, with HeLaS3 as a control, we assayed expression levels of Lin28a and Lin28b by Western blot, detected expression levels of let-7 family by QRT-PCR, and tested let-7 family activity by Asensors in HeLaS3/pAAV2neo-Lin28a and HeLaS3/pAAV2neo-Lin28b. Results demonstrated that both HeLaS3/pAAV2neo-Lin28a and HeLaS3/pAAV2neo-Lin28b could express Lin28a and Lin28b effectively. Compared with HeLaS3, the expression level of let-7 family except let-7e declined in HeLaS3/pAAV2neo-Lin28a. But declining extent among members of let-7 family was different. The let-7 family activity also decreased while the decreasing extent varied among members. Furthermore, the activity level was not consistent with its expression level for the same member in let-7 family. Compared with HeLaS3, both expression level and activity level of let-7 family in HeLaS3/ pAAV2neo-Lin28b were decreased. However, the decreasing extent of let-7 family expression changes was larger than that of HeLaS3/pAAV2neo-Lin28a while the decreasing extent of activity changes was similar. In this study, we established a method to detect and compare post-transcriptional inhibition level mediated by miRNA complementary targets. We firstly clarified the effect of Lin28a and Lin28b on let-7 family activity profile and found that this effect was not the same as that at expression level of let-7 family, suggesting that it was more comprehensive to understand miRNA regulation roles to detect both miRNA expression and activity. This paves a way for further research on mechanism of regulation of let-7 family.

Deletion of IV a2 gene from adenoviral genome by lambda-Red recombinase system and packaging of the recombinant adenovirus / 病毒学报

This investigation is to delete the most of the coding sequence (1104 bp) of the IV a2 gene in an adenovirus genome by a lambda-Red recombinase system-mediated PCR-targeting approach and rescue a recombinant adenovirus with IV a2 deletion. First, the template pAK of PCR targeting, containing kanamycin cassette, was constructed. Then, a linear fragment for PCR targeting, which had 39 bp homologous arms at both of its terminus, was amplified by PCR from the pAK. The pFG140 and the linear fragment were electroporated into E. coli BW25113/pIJ790 sequentially and the recombinant pFG140-deltaIV a2 (1104) was established by homologous recombination between the linear fragment and the pFG140 with aid of X-Red recombinase. The precise deletion of 1 104 bp fragment from IV a2 was confirmed by restriction endonucleases digestion and DNA sequencing. ORF of IV a2 was amplified by PCR from pFG140 and then cloned into the pAAV2neo vector. The recombinant adenovirus Ad5delta IV a2 (1104) was rescued by co-transfection of pFG140-deltaIV a2 (1104) and pAAV2neo-IV a2 into HEK293 cells. It was shown by Western Blot that IV a2 could not be detected in the Ad5deltaIV a2 (1104)- infected HEK293 cells. This study established a PCR-targeting strategy for manipulating adenovirus genome directly by a lambda-Red recombinase system, and a recombinant adenovirus with IV a2 deletion was obtained.

Study on severity, status of awareness and therapy regarding urinary incontinence in elderly people from rural areas in Jixian county,Tianjin / 中华流行病学杂志

Objective To understand the prevalence of urinary incontinence(UI)and its severity in rural elderly people,as well as to investigate the awareness on UI in the elderly and health-care service seeking behavior.Methods A cross-sectional study was carried out in two townships of Jixian county,Tianjin.A total of 743 people aged 60 years and over were selected under cluster sampling method.All the information was collected with a standardized structured questionnaire by face-to-face interview.Prevalence,severity,the awareness on UI in the elderly and their utilization of health-care service for its diagnosis and treatment were analyzed.Results Prevalence of UI was 33.38% among people aged 60 years and over in two townships of Jixian,higher in females than in males(43.15% vs.22.75%,χ~2=34.70,P＜0.0001).The prevalence rates of UI in 60-age group,65-age group,70-age group,75-age group,80-age group,85-95 age group were 28.64%,32.12%,34.08%,35.45%,47.76%,30.00%,respectively,and increased with age (for trend χ~2=219,P=0.029).Stress urinary incontinence(SUI)and urge urinary incontinence(UUI)were mild,while mixed urinary incontinence(MUI)were mainly moderate.The differences of severity of SUI,UUI,MUI between men and women did not show statistical significance(all P＞0.05).In 743 elderly people,more than half of the respondents had never heard of UI(50.20%,373/743)and only 170(22.88%)elderly people considered UI as a disease.630(84.79%)and 665(89.50%)elderly people in our research group did not know that such condition was curable and preventable.In 248 elderly people with UI,only 12(4.84%)of them ever seeking community health-care services in the health-care centers or hospitals.For the ones who did visit the centers,the purpose was only to seek for drug treatment.Conclusion Prevalence of UI appeared to be high among the elderly people in rural areas of Jixian county.Most of the elderly people were lack of knowledge about UI that hindered them from seeking for diagnosis and treatment in the clinics,plus the treatment program for UI was not standardized.Knowledge on UI and health care seeking behavior should be popularized and strengthened among elderly people living in the rural areas.

Study on the prevalence of urinary incontinence and its related factors among elderly in rural areas,Jixian county,Tianjin / 中华流行病学杂志

Objective To understand the prevalence of urinary incontinence(UI) and its related factors so as to develop a three-tier program for prevention of the disease.Methods A cross-sectional study was carried out in two townships of Jixian county,Tianjin,during July to November 2007.A total of 743 people aged 60 years and over were selected under cluster sampling,and all information were collected with a standardized structured questionnaire by face-to-face interview.All the data were analyzed with multivariate logistic regression method to explore the related factors for UI in the elderly.Results The overall prevalence of UI was 33.38 percent among people aged 60 years and over in two townships.Risk factors flor UI in men would include older age (OR=1.39).occupation(OR=5.00),awareness of UI(OR=1.91),having in chronic respiratory diseases(OR=2.23),prostate(OR=11.47),neurological(OR=11.76),or motor systems (OR=2.48).while protective factors would include high educational level(taking primary school or below as control group),OR for the junior middle school group appeared to be 0.35,for senior middle school group it was 0.77.and ofundergraduate group it was 0.53.Risk factors for UI in women would include older age(OR=1.31),constipation(OR=1.46),awareness of UI(OR=1.94),increased bodv nlass index(when normal weight group served as control group,OR in the overweight group was 1.03 and in the obesity group OR was 1.54),suffering from chronic respiratory diseases(OR=4.84).diabetes mellitus(OR=2.36),or motor system diseases(OR=1.37),more gravidity(OR=1.03),more parity(OR=1.02),suffering from perinea laceration(OR=1.72) and wound infection during delivery(OR=1.65),while protective factors would include physical exercises(OR=0.64).Conclusion Prevalence of UI was higher among the elderly people in rural areas of Jixian county,Tianjin.UI in the elderly might have been influenced by various factors which suggesting the intervention strategy should be targeted at those related factors as well as focusing on primary prevention.

To assess the demands of community-based health care service among 717 elderly / 中华流行病学杂志

<p><b>UNLABELLED</b>OBJECTIVE To understand the demands and influencing factors on the community-based health care service (CHS) among the elderly and to provide evidence of implementing health service for them</p><p><b>METHODS</b>Cluster sampling method was used on 717 persons aged 60 and above in Yanjiao Developing Area. A survey was carried out, using questionnaire and focus group discussion approaches.</p><p><b>RESULTS</b>Data showed that a two-week prevalence of diseases was 56.3% while the overall prevalence of chronic diseases was 92.1% with 70.0% of them suffering from more than two kinds of chronic diseases. The rates of ADL and IADL loss were 1.8% and 7.0% respectively with 37.0% of them sought medical consultation in the past two weeks. Major reason affecting the use of health service was low income. 57.7% of the elderly expressed their willingness of accepting CHS with the strongest demands as regular physical check-up, health education and consultation, specialized care services, home care service while major reasons hindering the acceptance of CHS could be referred to self-perception on good stamina and lack of awareness to CHS.</p><p><b>CONCLUSION</b>The demands of CHS in the elderly were greater than those in general population that called for CHS programs to be acceptable and relevant to meet the different needs.</p>

Evaluation on the effectiveness for self-management of hypertensive patients in a community / 中华流行病学杂志

<p><b>OBJECTIVE</b>To evaluate the acceptability of self-management project for patients with hypertension and the short term result, and provide the evidence of preventing and controlling hypertension using chronic disease self-management.</p><p><b>METHODS</b>Voluntary patients with hypertension were grouped into the control and experimental groups after matching age, sex, education levels and the number of chronic diseases between the two groups. Data collected through questionnaire after intervention were compared with the baseline data through analysis of covariance and chi(2) test.</p><p><b>RESULTS</b>Rate of awareness for the diagnostic criteria of hypertension, risk factors and associated diseases increased by 56.4%, 50.4% and 37.6% respectively, and the rate of high salt diet intake, body mass index and the times of visiting doctors decreased by 27.1%, 0.8 kg/m(2) and 2.7 times on average in experimental group, as compared with control group. The health status (energy, fatigue, shortness of breathe, pain) improved (P < 0.05). In the similar comparison, the rate in which the hypertensive patients took antihypertensive medicine increased by 20%, and the scores of cognitive symptoms management and of patient communication with physician increased by 3.9 and 1.1 points respectively. The rate of compliance to medication increased by 21.5% compared with themselves.</p><p><b>CONCLUSION</b>The awareness rate of hypertensive patients on knowledge and on unhealthy life style improved after implementing the self-management program. Their emotion, healthy behavior, health status and symptoms related to hypertension improved. Data showed that it was effective for hypertensive patients to use chronic diseases self-management.</p>